基于UPLC-MS分析技术测定不同产区制何首乌饮片中8个化学成分的含量

Determination of 8 chemical components in Polygonum multiflorum Thunb.from different production areas based on UPLC-MS

目的建立UPLC-MS分析技术同时测定不同产地制何首乌饮片中4种蒽醌类(大黄素甲醚、大黄素、大黄素甲醚-8-O-β-D-葡萄糖苷和大黄素-8-O-β-D-葡萄糖苷)及3种鞣质类(没食子酸、儿茶素、表儿茶素)和1种二苯乙烯类(反式二苯乙烯苷)活性成分的含量。方法采用Ultimate UPLC-XB C18(2.1 mm×50 mm,1.7μm)色谱柱,流动相A为0.1%甲酸水溶液,流动相B为乙腈,线性梯度洗脱,流速为0.2 mL·min^(-1);采用电喷雾离子源(ESI)、负离子扫描模式,在多反应监测(MRM)条件下进行定量分析;并用IBM SPSS Statistics 22软件对数据结果进行聚类分析。结果建立的UPLC-MS可以较好地分离8种成分,各成分在相应浓度范围内与峰面积线性关系良好;其定量限和检测限范围分别为0.89~13.00和0.20~4.00 ng·mL^(-1);精密度、重复性、稳定性的RSD值(n=5)均小于5%;加样回收率在99.47%~101.49%,RSD均小于2.5%;聚类分析结果显示12批制何首乌样品分为3类,其中S1、S2、S5、S8~S12归为Ⅰ类,S6、S7归为Ⅱ类,S3和S4归为Ⅲ类;第Ⅲ类样本(广西、广东)中3类成分的平均含量相对较高。结论该方法可靠、准确、快速、重复性好,可用于制何首乌的质量控制与品质相关研究,并为其临床安全用药提供依据。

Objective To determine 4 anthraquinones(physcion,emodin,physcion-8-O-β-Dglucoside and emodin-8-O-β-D-glucoside),3 tannins(gallic acid,catechin,and epicatechin)and 1 stilbene glycoside(trans-THSG)in Polygonum multiflorum decoction slices from different regions by UPLC-MS.Methods Ultimate UPLC-XB C18(2.1 mm×50 mm,1.7μm)column was used with 0.1%formic acid as mobile phase A and acetonitrile as mobile phase B.Linear gradient elution was performed at the flow rate of 0.2 mL·min^(-1).Quantitative analysis was performed with ESI source,negative ion scanning mode and multi-reaction monitoring(MRM).IBM SPSS Statistics 22 was used for cluster analysis.Results The UPLC-MS method well separated the 8 target components with good linearity in the corresponding concentration ranges.The limits of quantification and detection were 0.89~13.00 and 0.20~4.00 ng·mL^(-1),respectively.RSDs of precision,repeatability and stability(n=5)were all less than 5%.The recoveries ranged from 99.47%to 101.49%,and RSDs were all lower than 2.5%.Cluster analysis showed that the 12 batches of Polygonum multiflorum samples were divided into 3 categories,among which S1,S2,S5,S8~S12 were classified as classⅠ,S6 and S7 classified as classⅡ,while S3 and S4 classified as classⅢ.The average content of 3 components in classⅢsamples from Guangxi and Guangdong was relatively high.Conclusion The method is reliable,accurate,rapid and reproducible.It can be used to determine the content of 8 components in Polygonum multiflorum from different regions,and provide reference for the quality control of Polygonum multiflorum,as well as for safety and quality research.